Immunohistochemistry - Anti-Hsp90 alpha Rabbit mAb [65Y40N96]

Sample: Paraformaldehyde-fixed, paraffin embedded rat testis tissue

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.

Primary antibody: R015453 at 1:200 dilution

Secondary antibody: Goat Anti-Rabbit IgG (H+L), HRP conjugated at 1:1,000 dilution

DAB was used as the chromogen.

Counter stained with hematoxylin.

Positive/negative staining were presented.

Only the secondary antibody was used as the negative control.

Immunohistochemistry - Anti-Hsp90 alpha Rabbit mAb [65Y40N96]

Sample: Paraformaldehyde-fixed, paraffin embedded human lung cancer tissue

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.

Primary antibody: R015453 at 1:200 dilution

Secondary antibody: Goat Anti-Rabbit IgG (H+L), HRP conjugated at 1:1,000 dilution

DAB was used as the chromogen.

Counter stained with hematoxylin.

Positive/negative staining were presented.

Only the secondary antibody was used as the negative control.

Immunohistochemistry - Anti-Hsp90 alpha Rabbit mAb [65Y40N96]

Sample: Paraformaldehyde-fixed, paraffin embedded mouse liver tissue

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.

Primary antibody: R015453 at 1:200 dilution

Secondary antibody: Goat Anti-Rabbit IgG (H+L), HRP conjugated at 1:1,000 dilution

DAB was used as the chromogen.

Counter stained with hematoxylin.

Positive/negative staining were presented.

Only the secondary antibody was used as the negative control.

Immunofluorescence - Anti-Hsp90 alpha Rabbit mAb [65Y40N96]

Sample: HeLa cells

The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.5% Triton X-100 for 10 minutes and then blocked with 5% BSA in 0.1% PBS-Tween for 0.5 hours.

Primary antibodies: R015453 at 1:100 dilution and α-tubulin Mouse Monoclonal Antibody (Cat. No. LF209) at 1:100 dilution

Secondary antibodies: Goat anti-Rabbit (488) at 1:1,000 dilution (shown in green) and Goat anti-Mouse (555) at 1:1,000 dilution (shown in red)

Nuclei were stained with DAPI (shown in blue).

Western Blot - Anti-Hsp90 alpha Rabbit mAb [65Y40N96]

All lanes: R015453 at 1:1,000 dilution

Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates

Lane 2: HepG2 (Human hepatocarcinoma epithelial cell) whole cell lysates

Lane 3: HCT116 (Human colorectal carcinoma epithelial cell)  whole cell lysates

Lane 4: 293T (Human embryonic kidney cell) whole cell lysates

Lane 5: K562 (Human chronic myeloid leukemia cell) whole cell lysates

Lane 6: U87 (Human malignant glioblastoma epithelial cells) whole cell lysates

Lane 7:  Mouse heart whole tissue lysates

Lane 8: Mouse brain whole tissue lysates

Lane 9: PC-12 (Rat adrenal pheochromocytoma epithelial cell) whole cell lysates

Lane 10: Rat muscle whole tissue lysates

Lysates/proteins at 10 µg per lane.

Secondary antibody: Goat Anti-Rabbit IgG (H+L), HRP Conjugated (Cat. No. LF102) at 1:5,000 dilution

Predicted band size: 85 kDa

Observed band size: 85 kDa

Developed using the ECL technique (Cat. No. SQ201).